近幾年幹細胞領域的發展使得神經退化性疾病的治療透露出曙光，但是應用上有礙於神經幹細胞的稀少，希望尋求且有高產率及較好的神經分化效率的非神經細胞來源。這裡我們發現以人類胚胎幹細胞衍生前驅細胞(EMPs)可以經由Y-27632 的抑制有效地誘導神經型態出現。EMPs為從胚胎幹細胞所衍生的間葉前驅細胞，具備多分化潛能且不會形成畸胎瘤，也有免疫調節的作用，而且在組織培養環境下也較不易老化。另外在來源方面，EMPs可以無限的從胚胎幹細胞或是其他多功能性幹細胞，甚至於誘導多功能幹細胞來產生。我們從基因層面以及蛋白質層面來探討 EMPs經由Y-27632 誘導所產生的神經分化能力。在24小時內，Y-27632 可以快速地引發細胞延伸進而出現類神經型態，並且發現三個神經前驅因子Musashi1及neurogenin1的基因表現量上升，而GFAP則同時在基因及蛋白質表現上都發現有所提高。Y-27632可以有效抑制ROCKs的活性，其為控制細胞形狀有關的Rho家族的下游。因此，藉由Y-27632抑制ROCKs後，EMPs可以向類神經型態分化。我們以siRNA針對ROCKs做專一抑制或以抑制劑抑制ROCK II的下游myosin II所得結果與Y-27632類似，都可以使EMPs表現類神經型態。綜合以上結論，Rho/ROCKs 路徑的抑制可誘導EMPs產生類神經型態。

Current advances in stem cell biology have brought much hope for therapy of neuro-degenerative diseases. However, neural stem cells (NSCs) are rare adult stem cells, and the use of non-NSCs requires efficient and high-yielding lineage-specific differentiation prior to transplantation for efficacy. We report on the efficient differentiation of human embryonic stem cell-derived mesenchymal progenitors (EMPs) into a neural phenotype with use of Y-27632, a clinically compliant small molecular inhibitor of Rho kinase (ROCKs). EMPs are a population of mesenchymal progenitors derived from human embryonic stem cells (hESCs) which are capable of multilineage differentiation but do not form teratomas. These early mesenchymal progenitors are an ideal stem cell source since they are non-tumorigenic and immunomodulatory—similar to adult bone marrow mesenchymal stem cells—but do not easily senesce after in vitro culture, and are able to be regenerated indefinitely along from ESCs and other pluripotent stem cells including induced pluripotent stem cells. We treated EMPs with Y-27632 and investigated for differentiation capacity and neural lineages by gene expression, and protein expression. Y-27632 induced EMPs into a neural-like morphology, with rapid development of cell extensions and processes within 24 hours. Y-27632-treated EMPs express several neural progenitor marker included Musashi1and neurogenin1 at the RNA level and GFAP at RNA and protein level. EMPs can differentiate into a neural phenotype via inhibition of RhoA/ROCKs pathway by Y-27632. Moreover, both in specific knockdown Rho/ROCKs pathway or inhibited downstream of ROCK II have the same effects of neural-like morphology. Taken together, our data indicate that inhibition of RhoA-kinase induce of neural phenotype of embryonic stem cell-derived mesenchymal progenitors.

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